Primer Walking and Finishing

Primer walking refers to a process in which a Sanger sequencing primer is designed in a known DNA sequence adjacent to an unknown sequence resulting in a Sanger read that decodes the proximal unknown sequence as the read extends. Sequencing, designing a new oligo to the end of the newly decoded sequence and sequencing again iteratively resolves the unknown sequence across a gap or template in a directed, “walking” fashion.

Primer Walking

Beckman Coulter Genomics’ Primer Walking service is a stand alone service ideal for DNA templates too small for the practical application of a Next Generation technology but longer than the length of an average Sanger sequencing read. PCR^† amplicons, plasmid inserts, and entire plasmids are examples of templates regularly processed through Beckman Coulter Genomics’s Primer Walking service for single stranded or publication quality coverage.

Beckman Coulter Genomics’ Primer Walking service includes:

• Custom primer design and synthesis
• Single or double stranded sequencing coverage
• Sequence detection on ABI PRISM* 3730xl DNA sequencers
• Base calling and quality scoring
• Consensus sequence of assembled data

Genome Finishing

Genome Finishing uses the primer walking process to close gaps in a whole genome or large insert sequence. Finishing can also resolve repeat sequences a contig ordering when applied to a large or complex genome sequence project. Genome Finishing using primer walking can be incorporated into the strategy of any Whole Genome Sequencing projects initiated with Sanger or any of the Next Generation sequencing technologies.

2D Bar Coded Sample Tubes

Polypropylene tubes with integrated 2D bar code are available in packages of 96 tubes.